The promise of personalized medicine relies on the development of diagnostic tests to categorize patients within small cohorts who will benefit from specific treatments. 20/20 GeneSystems, Inc. (20/20) is a small business concern focused on the development of diagnostics for personalized medicine. Towards this end, 20/20 has developed, with the support of the NCI's SBIR program, an improved approach to immunohistochemistry ("Layered-IHC" also referred to as Layered Expression Scanning (LES)) that permits 10 or more biomarkers to be measured simultaneously from a single section of biopsied tissue while maintaining morphology. No other marketed technology has this dual capability. This SBIR Phase II Bridge Award proposal aims to develop, clinically validate and commercialize a series of companion diagnostic tests for cancer therapeutic agents, the efficacy of which may be influenced by the activity levels of proteins in the mTOR pathway (including PI3-K, Akt, and mTOR). The mTOR pathway is an important pathway in tumorigenesis and currently the target of several drugs on the market for kidney cancer (TORISEL(R) and AFINITOR(R)) as well as over a dozen compounds in Phase I-III trials for multiple tumor types. It has been demonstrated that a more complete characterization of the mTOR pathway is required to best determine the appropriateness of a specific therapy and as such measurement of a panel of multiple biomarkers will be required. Thus 20/20's L-IHC system is a most appropriate system for the development of mTOR companion diagnostics. This SBIR Phase II Bridge Award proposal seeks to develop what is likely the first mTOR companion diagnostic, PredicTOR", and clinical validation of this test in two or three cancer types, specifically kidney cancer, breast cancer and lung cancer. This work will be done in consultation with the FDA and will include the necessary protocols and procedures to obtain regulatory acquiescence or approval of the products as laboratory developed tests (LDTs). To increase the likelihood of successful and timely commercialization, applicant plans to match the requested $2.75 million in government funding (over 3 years) with at least $3.75 million in private sector investment. This private investment will come from one or more strategic partners ($2 million) and individual investors ($1.75 million). 20/20 has a proven track record in procuring both equity investors and corporate partnerships and has already successfully commercialized innovative products developed with federal funding. Commercialization of these products is expected by the conclusion of the funding period. PUBLIC HEALTH RELEVANCE: This SBIR Phase II Bridge Award proposal seeks to develop what is likely the first mTOR companion diagnostic, PredicTOR", and clinical validation of this test in two or three cancer types. This work will utilize 20/20 GeneSystems, Inc. proprietary L-IHC t=technology and will be done in consultation with the FDA and will include the necessary protocols and procedures to obtain regulatory acquiescence or approval of the products as laboratory developed tests (LDTs).